Fig 2: Knockdown of LINC00483 decreases tumor growth in gastric cancer xenograft model. a Tumor volume was measured every week. b Tumor weight was detected at the end. c qRT-PCR assay detected the expression levels of LINC00483, miR-490-3p and MAPK1 in tumor tissues. d Western blot assay examined the protein levels of MAPK1, c-Myc, Bax and MMP-9 in tumor tissues. sh-LINC00483: xenograft tumor formed using MGC-803 cells with LINC00483 knockdown using shRNA; sh-NC: xenograft tumor formed using MGC-803 cells with transfection of shRNA negative control. *P < 0.05 compared with sh-NC group

Fig 3: The ERK1/2 pathway controls CSC phenotypes in CD44(+) HNSCC cells.a Phospho-RTK protein array analysis using CD44(+) and CD44(-) cells. b Western blotting of CD44, Nanog, phosphorylated ERK1/2, total ERK2, phosphorylated JNK1/2, total JNK2, phosphorylated p38, total p38, and ß-actin in CD44(+) and CD44(-) cells. c Western blotting of total ERK1/2, total ERK1, total ERK2, CD44, and Nanog in CD44(+) cells transduced with sh.ERK1/2 or sh.Scr. d IF of CD44(+) cells transduced with sh.ERK1/2 or sh.Scr for CD44 and Nanog. e Single cell assay and f colony formation assay of CD44(+) SCC-15 and QLL-1 cells following transduction with sh.ERK1/2 or sh.Scr. g Migration and Invasion assay of CD44(+) SCC-15, SCC-25, and QLL-1 cells following transduction with sh.ERK1/2 or sh.Scr. h Western blotting of ERK1/2, N-cadherin, Snail, Slug, Zeb1, and ß-actin and immunofluorescent images of N-cadherin and Snail staining in CD44(+) cells transduced with sh.ERK1/2 or sh.Scr. Scale bar = 20 µm. Error bars represent standard deviation. *p < 0.05.

Fig 4: Effects of miR-145 on SW480 cells via MAPK1. (A) Identification of MAPK1 as a potential target of miR-145. (B) Expression of MAPK1 in normal and colon adenocarcinoma cell lines. (C) Expression of MAPK1 in miR-NC- and miR-145 mimics-transfected SW480 cells. (D) Luciferase assay. (E) Expression of MAPK1 in si-NC- or si-MAPK1-transfected SW480 cells. Effect of the suppression of MAPK1 on SW480 cell (F) proliferation, (G) colony formation and (H) apoptosis. The values are expressed as the mean ± standard deviation of three biological replicates (*P<0.05). miR, microRNA; NC, negative control; MAPK1, mitogen-activated protein kinase 1; si-, small interfering RNA; WT, wild-type; MUT, mutant (magnification, ×200).

Fig 5: The expression levels of LINC00483 and MAPK1 are up-regulated in gastric cancer. a, b qRT-PCR assay detected the levels of LINC00483 and MAPK1 in gastric cancer tissues and normal samples. n = 30. c Western blot assay was performed to measure the MAPK1 protein level in gastric cancer tissues and normal tissues. d The association between levels of LINC00483 and MAPK1 in gastric cancer tissues was assessed. e–g The expression levels of LINC00483 and MAPK1 were detected in gastric cancer cells via qRT-PCR or western blot. GC: gastric cancer. *P < 0.05 compared with normal or GES-1 group